Chemical Variability and Antioxidant Activities of the Essential Oils of the Aerial Parts of Ammoides verticillata and the Roots of Carthamus caeruleus and their Synergistic Effect in Combination.

AIM AND OBJECTIVE: Oxidative stress is implicated in the development and progression of many diseases. Some of the appropriate actions that could taken to resolve the problem of these diseases are search for new antioxidant substances isolated from plants. The aims of this study were to research the intraspecies variations of A. verticillata and C. caeruleus essential oils from 8 locations using statistical analysis, the in vitro antioxidant properties of collective essential oils and in combinations. MATERIALS AND METHODS: The essential oils were analyzed by GC and GC-MS. The intraspecies variations of the essential oil compositions were discussed using principal component analysis (PCA) and cluster analysis (CA). The antioxidant properties were evaluated DPPH-radical scavenging activity and ß-carotene bleaching test. RESULTS: The main components of Ammoides verticillata collective essential oil (Coll EO) were thymol (30.5%), carvacrol (23.2%), p-cymene (13.1%), limonene (12.5%) and terpinene-4-ol (12.3%). While roots of Carthamus caeruleus essential oil were dominated by carline oxide (86.2%). The chemical variability allowed the discrimination of two main Groups for both Coll EOs. A direct correlation between the altitudes, climate and the chemical compositions was evidenced. Ammoides verticulata and Carthamus caeruleus Coll Eos showed good antioxidant activity. In binary mixture, the interaction between both Coll Eos and between oils rich of thymol and/or carvacrol with carlina oxide produced the best synergistic effects compared to individual essential oils and the synthetic antioxidant (BHT). CONCLUSION: Ammoides verticillata and Carthamus caeruleus essential oil blends can be used as a natural food preservative and alternative to chemical antioxidants.

Hemisyntheses and In-silico Study of New Analogues of Carlina Oxide from Carthamus Caeruleus Roots.

AIM AND OBJECTIVE: Nowadays, developing effective antibiotics for bacterial control has become difficult due to increased resistance to the available medicines in the market. Essential oils possess interesting biological properties as some of their components have very powerful antiviral and antibacterial properties. Carthamus caeruleus is a plant that has antibacterial and antioxidant activity due to the presence of an acetylenic compound, Carlina oxide. The aim of this work was to provide, for the first time, the chemical modifications to the structure of Carlina oxide and the insilico study of these analogues. MATERIALS AND METHODS: The essential oil of Carthamus caeruleus was extracted by steam distillation in a Clevenger-type apparatus. Carlina oxide component was separated by column chromatography. Five new analogues were synthetized and identified by spectroscopic analyses (RMN, IR and SM). Molecular docking simulation study was performed using Molecular Operating Environment software (MOE) on three enzymes of bacterial origin (Streptococcus pyogenesis and Enterococcus faecalis). RESULTS: Five new compounds derived from Carlina oxide were synthesized (IM8-IM12), and their structures were characterized by infrared (IR), 1H and 13C nuclear magnetic resonance (NMR). The new synthesized compounds were evaluated as mSpeB, DHFR from Enterococcus faecalis and DNA gyrase inhibitors by a docking analysis using MOE. These results show interesting ligand interactions with the three enzymes, and the best result was attributed to the complexes formed with IM9, which had the lowest score. CONCLUSION: In fact, these new compounds could lead to powerful approaches for the research and development of new antibiotics.

Dehydroabietylamine, A Diterpene from Carthamus tinctorious L. Showing Antibacterial and Anthelmintic Effects with Computational Evidence.

BACKGROUND: Plant-based drugs provide an outstanding contribution to modern therapeutics, and it is well known that the presence of different phytochemicals is responsible for such pharmacological effects. Carthamus tinctorius L. is one such medicinally important plant whose different solvent extracts have been reported with several pharmacological effects like antibacterial, hepatoprotective, and wound healing. The exploration of phytoconstituents from such a medicinally important plant for different pharmacological effects could produce new and effective drugs to treat human diseases. OBJECTIVE: The present study attempts to explore the antibacterial and anthelmintic properties of dehydroabietylamine, a diterpene isolated from Carthamus tinctorius L. followed by the in silico elucidation of its probable mode of action. METHODS: The minimum inhibition concentration (MIC) and minimum bactericidal concentration (MBC) of dehydroabietylamine were assessed against Staphylococcus aureus and Pseudomonas aeruginosa, using micro- broth dilution method. The anthelmintic activity of was determined to assess the time taken for paralysis and death of Pheretima Posthuma at different concentrations. Additionally, molecular docking study was conducted to understand the interaction between dehydroabietylamine with target proteins identified for both antibacterial and anthelmintic activity viz., glucosamine-6-phosphate synthase and ß-Tubulin, respectively. RESULTS: The dehydroabietylamine showed the significant MIC for S. aureus (12.5 µg/ml) and P. aeruginosa (6.25µg/ml), respectively. The result of anthelmintics effect of dehydroabietylamine was found to be dosedependent and compared to the standard drug, albendazole. CONCLUSION: The interactions of dehydroabietylamine with the two target proteins with high binding affinity indicated the probable inhibition of target proteins, which could be the cause for prominent antibacterial and antihelminthic effects.

[Nature-effect relationship research of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos based on nature combination].

Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos are commonly traditional Chinese medicines with invigorating blood circulation and eliminating blood stasis,but they are different in effects due to differences in five tastes and four properties. In this study,Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos are selected as research vectors to obtain the active ingredients and targets through TCMD,TCMSP database and Ch EMBL database. The protein interaction information of the target is used to obtain from STRING online database,then imported into Cytoscape software to plot the protein interaction network and perform GO enrichment analysis.The results show that the heart-liver protein interaction network,involving blood circulation and hepatic lipid metabolism,thereby exerting the effect of activating blood circulation. The cold-bitter protein interaction network involves the biological process of vasoconstriction,thereby exerting cooling blood and the efficacy of eliminating phlegm. The warm-pungent protein interaction network involves blood coagulation,lipid metabolism and other biological processes to play the role of phlegm pain. Through analysis,it is found that the relationship between pharmacological efficacy and medicinal properties has a certain degree of specificity,which facilitates the subsequent scientific and systematic study of medicinal properties on the basis of this study.

Extracts from the Mediterranean Food Plants Carthamus lanatus, Cichorium intybus, and Cichorium spinosum Enhanced GSH Levels and Increased Nrf2 Expression in Human Endothelial Cells.

The Mediterranean diet is considered to prevent several diseases. In the present study, the antioxidant properties of six extracts from Mediterranean plant foods were assessed. The extracts' chemical composition analysis showed that the total polyphenolic content ranged from 56 to 408 GAE mg/g dw of extract. The major polyphenols identified in the extracts were quercetin, luteolin, caftaric acid, caffeoylquinic acid isomers, and cichoric acid. The extracts showed in vitro high scavenging potency against ABTS•+ and O2 •- radicals and reducing power activity. Also, the extracts inhibited peroxyl radical-induced cleavage of DNA plasmids. The three most potent extracts, Cichorium intybus, Carthamus lanatus, and Cichorium spinosum, inhibited OH•-induced mutations in Salmonella typhimurium TA102 cells. Moreover, C. intybus, C. lanatus, and C. spinosum extracts increased the antioxidant molecule glutathione (GSH) by 33.4, 21.5, and 10.5% at 50 µg/ml, respectively, in human endothelial EA.hy926 cells. C. intybus extract was also shown to induce in endothelial cells the transcriptional expression of Nrf2 (the major transcription factor of antioxidant genes), as well as of antioxidant genes GCLC, GSR, NQO1, and HMOX1. In conclusion, the results suggested that extracts from edible plants may prevent diseases associated especially with endothelium damage.

Gas chromatography coupled to mass spectrometry characterization, anti-inflammatory effect, wound-healing potential, and hair growth-promoting activity of Algerian Carthamus caeruleus L (Asteraceae).

OBJECTIVES: The roots of Carthamus caeruleus have been used by the population of Northern Algeria to treat several pathological conditions, including wound healing and hair growth. The present study was conducted to evaluate the anti-inflammatory activity, wound-healing potential, and hair growth-promoting activity attributed to C. caeruleus root. MATERIALS AND METHODS: In this study, we have investigated the anti-inflammatory effect using carrageenan-induced paw edema test, evaluated the wound-healing potential by linear incision wound model, and evaluated hair growth activity using in vivo hair growth-promoting test attributed to C. caeruleus root. Preliminary phytochemical screening and gas chromatography coupled to mass spectrometry (GC/MS) characterization were also performed. RESULTS: It was found that the methanolic extract of C. caeruleus was characterized by the presence of tannins, flavonoids, anthocyanins, leucoanthocyanins, sennosides, free quinones, saponins, glycosides, mucilage, and coumarins. The GC/MS analysis could identify 22 compounds and showed that the major chemical constituents were palmitic acid (12.88%), mono(2-ethylhexyl) phthalate (12.75%), and 5-(hydroxymethyl)-2-furancarboxaldehyde, (9.19%). The phytoextract strongly inhibited (P < 0.001) paw edema formation in mice. The roots of C. caeruleus also showed a significant (P < 0.05) wound-healing and hair growth-promoting effects. CONCLUSION: The results indicate the richness of the roots of the Algerian C. caeruleus in biomolecules. These molecules exhibit an excellent reducing inflammation activity, a wound-healing property, and an interesting hair-promoting growth activity. All in all, the findings promote the usage of the Algerian C. caeruleus as an effective and a safe potential skincare alternative remedy.

Seed Polyphenolic Profile, Antioxidative Activity, and Fatty Acids Composition of Wild and Cultivated Carthamus Species.

Total flavonoid content (TFC) and cyanidin-3-glucoside (Cyd-3-glu) of seed and seed coat extract of 16 genotypes from five species of Carthamus were studied, and their major polyphenolic compounds and antioxidant activity of the seed coat extracts were determined using HPLC analysis and DPPH assay, respectively. Additionally, fatty acids composition of the seed oil was analyzed by GC. In general, TFC and Cyd-3-glu content of seed coat extracts were higher than those of seed extracts. A novel breeding line with black seed coat (named A82) depicted lower TFC (3.79 mg QUE/g DW) but higher Cyd-3-glu (24.64 mg/g DW) compared to the white and other seed-pigmented genotypes. DPPH radical scavenging activity showed a strong association with Cyd-3-glu content (r = 0.84), but no correlation with TFC (r = -0.32). HPLC analysis of seed coat extracts revealed that four compounds were dominant constituents including rutin (7.23 - 117.95 mg/100 g DW), apigenin (4.37 - 64.88 mg/100 g DW), quercetin (3.09 - 14.10 mg/100 g DW), and ferulic acid (4.49 - 30.41 mg/100 g DW). Interestingly, the genotype A82 with an appropriate polyunsaturated/saturated fatty acids index (5.46%) and a moderate linoleic fatty acid content (64.70%) had higher nutritional and pharmaceutical value than all the other genotypes.

Cardioprotective Effect of Hydroxysafflor Yellow A via the Cardiac Permeability Transition Pore.

Myocardial ischemia damages cardiac myocytes in part via opening of the mitochondrial permeability transition pore. Preventing this pore's opening is therefore a useful therapeutic goal in treating cardiovascular disease. Hydroxysafflor yellow A has been proposed as a nontoxic alternative to other agents that modulate mitochondrial permeability transition pore opening. In this study, we proposed that hydroxysafflor yellow A prevents mitochondrial permeability transition pore formation in anoxic cardiac myocytes, and thus protects the cell from damage seen during reoxygenation of the cardiac myocytes. Experiments with hydroxysafflor yellow A transport in aerobic myocytes show that roughly 50% of the extracellular dye concentration crosses the cell membrane in a 2-h incubation. In our anoxia/reoxygenation protocol, hydroxysafflor yellow A modulated both the reduction of viability and the loss of rod-shaped cells that attend anoxia and reoxygenation. Hydroxysafflor yellow A's protective effect was similar to that of cyclosporin A, an agent known to inhibit mitochondrial permeability transition pore opening. In additional experiments, plated myocytes were loaded with calcein/MitoTracker Red, then examined for intracellular dye distribution/morphology after anoxia/reoxygenation. Hydroxysafflor yellow A-containing cells showed a cardioprotective pattern similar to that of cyclosporin A (an agent known to close the mitochondrial permeability transition pore). We conclude that hydroxysafflor yellow A can enter the cardiac myocyte and is able to modulate anoxia/reoxygenation-induced damage by interacting with the mitochondrial permeability transition pore.

Molecular dynamics simulation integrating the inhibition kinetics of hydroxysafflor yellow A on α-glucosidase.

Inhibition of α-glucosidase has attracted the attention of researchers due to its connection to type-2 diabetes. Hydroxysafflor yellow A (HSYA) extracted from Carthamus tinctorius L. is a natural antioxidant used in traditional Chinese medicine. In this study, the effect of HSYA on α-glucosidase was evaluated using inhibitory kinetics based on the antioxidant properties of HSYA and by performing computational simulation integration methods. HSYA reversibly inhibited α-glucosidase in a competitive inhibition manner and the evaluated kinetic parameters were IC50 = 1.1 ± 0.22 mM and Ki = 1.04 ± 0.23 mM, respectively. The results of spectrofluorimetry showed that the inner hydrophobic regions of α-glucosidase, which are mostly in the active site, were exposed to the surface with increasing HSYA concentrations, indicating that the inactivation of α-glucosidase by HSYA was accompanied by regional unfolding. The molecular dynamics simulations indicated that the four rings of HSYA interact with four residues such as G217, A278, H279, and G280 at the entrance of the active site. Our study provides insight into the inhibition of α-glucosidase and the accompanying structural changes by HSYA. Based on its α-glucosidase-inhibiting effect and its potential as a natural antioxidant, HSYA is a potential agent for treating α-glucosidase-associated type-2 diabetes.

New lipoxygenase and cholinesterase inhibitory sphingolipids from Carthamus oxyacantha.

Two new sphingolipids: oxyacanthin A [(2S,3S,4R)-2-{[(2R,5E)-2-hydroxyoctadec-5-enoyl]amino}hexaeicosane-1,3,4-triol; 1] and B [(2S,3S,4R)-2-{[(2R,5E)-2-hydroxyoctadec-5-enoyl]amino}hexaeicosane-1,3,4-triol-1-O-ß-D-glucopyranoside; 2], together with 1-octacosanol, ß-sitosterol, ß-sitosterol 3-O-ß-D-glucopyranoside and luteolin 7-O-ß-glucopyranoside were isolated from the methanolic extract of the whole plant of Carthamus oxyacantha. Their structures were elucidated using (1)H and (13)C NMR spectra and 2D NMR analyses (HMQC, HMBC and COSY) in combination with mass spectrometry (EI-MS, HR-EI-MS, FAB-MS and HR-FAB-MS) experiments and in comparison with the literature data of the related compounds. Both the compounds 1 and 2 showed inhibitory potential against lipoxygenase (LOX) in a concentration-dependent manner with IC50 values 83.3 ± 1.3 and 245.7 ± 1.1 µM, whereas compound 2 showed inhibition against enzymes acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) with IC50 values 65.3 ± 0.1 and 93.6 ± 0.1 µM, respectively.

[Data mining analysis of regularity of formulas containing Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos medicin pair in Dictionary of Chinese Medicine Prescription].

In this study, formulas containing Salviae Miltiorrhizae Radix et Rhizoma-Carthami Flos in the database of Dictionary of Chinese Medicine Prescription (DCMP) were extracted by using traditional Chinese medicine inheritance support system (TCMISS). The drugs pairs and formula composition rules were analyzed with data mining methods, such as association rules, improved mutual information method and complex system entropy clustering. Totally 39 formulas were included in this study and involved 280 Chinese medicines. The top 5 Chinese medicines most frequently used were Danggui (Angelica sinensis), Chuanxiong (Ligusticum chuanxiong), Xiangfu (Cyperi Rhizoma), Baishao(Radix Paeoniae Alba), Taoren(Prunus persica) and Shengdihuang (Radix Rehmanniae Recens). Six core medicinal pairs were obtained through clustering analysis, namely Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Xiangfu (Cyperi Rhizoma)-Honghua (Carthami Flos), Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Baishao (Radix Paeoniae Alba)-Honghua (Carthami Flos), Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Danggui (A. sinensis)-Xiagnfu (Cyperi Rhizoma)-Honghua (Carthami Flos), Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Danggui (A. sinensis)-Baishao (Radix Paeoniae Alba)-Honghua (Carthami Flos), Honghua (Carthami Flos)-Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Baishao (Radix Paeoniae Alba)-Danggui (A. sinensis), Danshen (Salviae Miltiorrhizae Radix et Rhizoma)-Baishao (Radix Paeoniae Alba)-Honghua (Carthami Flos)-Danggui (A. sinensis). The support degree was set at 11 (38.46%), with a confidence coefficient of 80%, and then 38 associated pairs were screened. These results suggested that Salviae Miltiorrhizae Radix et Rhizoma, Carthami Flos is often combined with herbs for activating blood and promoting circulation of qi to treat gynecopathy, stasis blood pain syndrome, stroke and other syndromes.

Effect of Safflower Oil on Concentration of Conjugated Linoleic Acid of Kefir Prepared by Low-fat Milk.

BACKGROUND: Conjugated linoleic acid (CLA) is a special fatty acid in dairy products with unique antioxidant and anti-cancerous effects. Kefir, a milk product, comprises normalized homogenized cow's milk, the fructose and lactulose syrup as well as a symbiotic starter which has improved probiotic characteristics. The study was aimed to discuss patents and to examine the effect of different safflower oil concentrations on CLA content of the kefir drink prepared by low-fat milk. MATERIAL AND METHODS: Safflower oil was added at 0.1, 0.3 and 0.5% (V/V) to low-fat cow's milk and six formulations of kefir samples were prepared. The CLA content of the kefir products was measured at pH=6.0 and pH=6.8 by gas chromatography. Acid and bile tolerance of bacterial microenvironment in the products were also determined. RESULTS: Substitution of natural fat content of milk with safflower oil resulted in proportional increase in the CLA contents of kefir in a dose dependent manner. The highest concentration of CLA was found under 0.5% (V/V) of safflower oil at pH 6.0 and temperature of 37 °C. Adding the Safflower oil into milk used for kefir production, increased CLA content from 0.123 (g/100 g) in pure safflower free samples to 0.322 (g/100 g) in samples with 0.5% (V/V) of safflower oil. CONCLUSION: The current study revealed that substitution of safflower oil with natural fat of cow's milk may help the production of kefir samples with remarkable increase in CLA content of final product.

Physical characterization of wax/oil crystalline networks.

The objective of this research was to evaluate the physical properties of different types of wax/oil systems. Olive (OO), corn (CO), soybean (SBO), sunflower (SFO), safflower (SAFO), and canola (CAO) oils were mixed with sunflower oil wax (SFOW), paraffin wax (PW), and beeswax (BW) at different concentrations (1% to 10%). Results from this study show that the physical properties of wax/oil systems is affected not only by the concentration and type of wax used, but also by the type of oil used. In general, wax/oil systems formulated with SFOW generated crystalline networks with high enthalpies (1 to 22 J/g) and high G' values (2 to 6 × 10(6) Pa) compared with the values obtained for BW and PW. SFOW crystalline networks were characterized by needle-like crystals independently of the wax concentrations and type of oil used. BW crystalline networks, however, were characterized by different crystal morphologies (needle-like or spherulites) depending on the wax concentration and type of oil used. PW samples were characterized by a crystalline network formed by needle- and platelet-like crystals. Enthalpy values of BW and PW samples were similar (0.3 to 20 J/g), but BW samples resulted in significantly higher (P < 0.05) G' values in the 5% and 10% samples with values of 3.9 × 10(6) and 6.1 × 10(5) Pa for 10% BW and PW, respectively.

[Study of screening nephroprotective bioactive substances based on triple-color fluorescence probes in Carthami flos].

In this study, an approach based on triple-color fluorescence probes was developed for screening potential nephro-protective bioactive substances. Three fluorescent probes (i. e. FDA, MTR and Hoechst 33342) were used to label HK-2 cells injured by doxorubicin hydrochloride, and cellular fluorescence images were subsequently acquired and analyzed by a cellular-fluorescence image microscopy platform. The established method was applied to screening 53 components of Carthami Flos, and three components C17, C18 and C19 were found to exhibit nephroprotective effects against doxorubicin hydrochloride induced injury on HK-2 cells. Eight compounds (i. e. hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside, 6-hydroxykaempferol-3,6-di-O-gluco-side or 6-hydroxykaempferol-6, 7-di-O-glucoside, 6-hydroxykaempferol-3-O-rutinoside, 6-hydroxykaempferol-3-O-glucoside or 6-hydroxykaempferol-7-O-glucoside, rutin, isoquercetin, and kaempferol-3-O-rutinoside) in components C17, C18 and C19 were preliminarily identified by liquid chromatography-mass spectrometry (LC-MS). Isoquercetin, rutin, kaempferol-3-O-rutinoside, and hydroxysafflor yellow A were confirmed by comparing with reference substances, Further study indicated that these four compounds had moderate nephroprotective effects, while isoquercetin showed a significant nephroprotective effect in a dose-dependent manner. These results suggest that isoquercetin, rutin, kaempferol-3-O-rutinoside and hydroxysafflor yellow A might be the nephroprotective bioactive substances in Carthami Flos.

[Analysis on composition regularity of carthami flos-containing traditional Chinese medicines].

To analyze the composition regularity of Carthami Flos-containing prescriptions of the Drug Standards of Ministry of Health of People's Republic of China-Traditional Chinese Medicine Preparations (the ministerial standards for Traditional Chinese Medicine) based on the traditional Chinese medicine inheritance support system (TCMISS, RZDZ No. 0389952). Efforts were made to identify 331 prescriptions containing Carthami Flos and summarize 16 attending functions and 10 commonly used drug combinations. Three commonly used drug combinations were selected for an in-depth analysis on Carthami Flos's combined administration regularity. Based on Carthami Flos's attending functions, its effects in paralysis, traumatic injuries and dysmenorrheal were compared to analyze Carthami Flos's core drug combinations for treating different diseases. The regularity of clinical administration and the characteristics of commonly used drug combinations were summarized to provide reference for Carthami Flos's clinical application and new ideas for new drug R&D. Carthami Flos prescriptions was mainly used to treat blood stasis and pain and mostly combined with drugs that could activate blood, promote the circulation of qi and dispel pathogenic wind to treat Qi-stagnation and blood stasis caused by various pathogenic factors such as wind, cold and dampness.

Carthamus, Salvia and Stachys species protect neuronal cells against oxidative stress-induced apoptosis.

CONTEXT: Finding effective therapies for neurodegenerative diseases is of utmost importance for the aging population. Plants growing in Iran are rich sources of antioxidants and active phytochemicals. OBJECTIVE: The protective capacity of plants, with a special focus on those with reported antioxidant or neuroprotective potential or nervous system-related applications in folk medicine, was tested against oxidative stress-induced apoptosis. MATERIALS AND METHODS: Aerial parts of 20 plants including Carthamus, Salvia, and Stachys species were extracted with 80% methanol and dichloromethane and preincubated with neuronal PC12 cells for 3 h. Oxidative stress and apoptosis were induced by hydrogen peroxide (75 µM, 1 h exposure). Cell viability and intracellular reactive oxygen species (ROS) were measured by MTT and 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assays, respectively, while apoptosis was determined by annexin V-FITC/propidium iodide staining by a flow cytometer. RESULTS: Eighty percent methanol extracts of Carthamus oxyacantha Bieb. (Asteraceae), Salvia santolinifolia Boiss. (Lamiaceae), and Salvia sclarea L. (Lamiaceae) at the concentration of 100 µg/ml showed significant neuroprotection in the MTT assay by 38.7, 34.7, and 39.5%, respectively, and inhibited intracellular ROS by 48.6, 61.9, and 61.4%, respectively. The first two extracts also significantly inhibited apoptosis. Dichloromethane extracts of C. oxyacantha and Stachys pilifera Benth. (Lamiaceae) at the concentration of 25 µg/ml showed neuroprotection by 27.5 and 26.5%, respectively, and inhibited ROS by 44.5 and 39.4%, respectively. CONCLUSION: The above-mentioned plants seem to have important biological activities and their further study may lead to the discovery of new natural therapeutics useful against disorders such as Alzheimer and Parkinson diseases.

Pro-angiogenic effects of Carthami Flos whole extract in human microvascular endothelial cells in vitro and in zebrafish in vivo.

AIM: Carthami Flos (CF) is a Chinese herb traditionally used for cardiovascular disease and bone injury in China with pharmacological effects on improving blood circulation. The aim of this study was to investigate the angiogenic potential of CF whole extract (extracted by boiling with water, followed by ethanol) and the underlying mechanisms in human microvascular endothelial cells (HMEC-1) in vitro and in transgenic TG(fli1:EGFP)(y1)/+(AB) zebrafish with transgenic endothelial cells expressing EGFP (Enhanced Green Fluorescent Protein) in vivo. METHODS: Effects of CF whole extract on cell proliferation, migration and tube formation in HMEC-1 cells in vitro were detected by MTT assay, wound healing assay and tube formation assay. Its angiogenic effect in zebrafish was investigated by monitoring the sprout number in the sub-intestinal vessel (SIV), and the underlying mechanisms were tested by quantitative real-time PCR. RESULTS: CF whole extract increased cell proliferation, migration and tube formation in vitro in HMEC-1 cells. Its angiogenic effect was also confirmed in vivo in zebrafish by increasing the sprout number in the SIV. As determined by quantitative real-time PCR, CF whole extract up-regulated the expression of angiogenesis-related genes in zebrafish, including angiogenic and its associated growth factors and receptors (e.g. IGF1, CTGF, NRP2, and VEGFR3), transcription factor (e.g. HIF1A), matrix degradation and endothelial cell migration-related factors (e.g. MMP2, MMP9, TIMP2, PLG and PLAU), cell adhesion molecules (e.g. ITGAV, ITGB3, beta-catenin and PECAM1), tubule formation factors (e.g. ANGPT1, TIE-2, PDGFR-B, CDH5, S1PR1, FGF2, Shh, and TGFRB1), and blood vessel maturation/formation factor (e.g. Ephrin B2). CONCLUSIONS: CF whole extract increased angiogenesis in HMEC-1 cells in vitro and in zebrafish in vivo with multiple mechanisms.

Evaluation of antibacterial, antifungal, and antioxidant activities of safflower natural dyes during flowering.

Two Carthamus tinctorius varieties (Jawhara and 104) were studied in order to investigate their natural dyes contents and biological activities. Obtained results showed that quinochalcone contents and antioxidant activities varied considerably as function of flowering stages. So flowers at fructification stage contained the highest carthamin content with the strongest antioxidant capacity with all assays (FRAP, DPPH, and chelating power methods). In parallel, we showed a decrease in the content of precarthamin. The quantitative variation of these molecules could be due to colour change of C. tinctorius flowers. Correlation analysis indicated that the ABTS method showed the highest correlation coefficients with carthamin and precarthamin contents, that is, 0.886 and 0.973, respectively. Concerning the regional effect, the contents of precarthamin and carthamin varied significantly (P < 0.05) at studied regions with the optimum production given by samples of Beja (902.41 µg/g DW and 42.05 µg/g DW, respectively, at flowering stage). During flowering, the antimicrobial activity of these two natural dyes increased where the maximum inhibitory effect mentioned with carthamin mainly against E. coli (iz = 25.89 mm) at fructification stage. Therefore, the increased frequency of resistance to commonly used antibiotics leads to the search for new effective natural drugs at food and pharmaceutical industries.

Effect of honghua (Flos Carthami) on nitric oxide production in RAW 264.7 cells and alpha-glucosidase activity.

OBJECTIVE: To study the effects of extracts from Honghua (Flos Carthami) on lipopolysaccharide induced nitric oxide (NO) production in RAW 264.7 cells and the influence of the extracts on yeast a-glucosidase activity. The total flavonoid content of the extracts was also determined. METHODS: Cytotoxicity of the extracts to RAW 264.7 cells was evaluated by the ATPlite method. Inhibitory effects of the extracts on NO production were evaluated by Griess assay. Curcumin was used as a positive control. Screening of extracts for potential a-glucosidase inhibitors was done by a fluorometric assay. The assay was based on the hydrolysis of 4-methylumbelliferyl-a-D-glucopyranoside to form the fluorescent product, 4-methylumbelliferone. Acarbose was used as a positive control. The total flavonoid content was tested using kaempferol as the standard. RESULTS: There were significant inhibitory effects on NO production when the extracts were 25-100 microg/ mL (P < 0.05) and curcumin was 2-4 microg/mL (P < 0.001). The extracts showed an inhibitory effect on alpha-glucosidase activity at the concentrations of 15.6-125 microg/mL with a half maximal (50%) inhibitory concentration (IC50) of (32.8 +/- 5.7) microg/mL, compared with the IC50 of acarbose at (1.8 +/- 0.4) microg/mL. There was a significant difference between the two IC50 values (P < 0.001). The total content of flavonoids per gram of dried herb was 1.14 mg. CONCLUSION: Honghua (Flos Carthami) showed inhibitory effects on NO production in activated RAW 264.7 macrophage cells and an inhibitory effect on yeast alpha-glucosidase. There might be a relationship between these pharmacological effects and its flavonoid content.

The central analgesic and anti-inflammatory activities of the methanolic extract of Carthamus oxycantha.

The plant extract and fractions of Carthamus oxycantha (Compositae) were assessed for analgesic and antiinflammatory activities. Acetic acid and formalin-induced nociception, hot plate and carrageenan-induced rat paw oedema tests were employed to evaluate the analgesic and anti-inflammatory potential of the plant extract. The intraperitoneal (i.p.) administration of the methanolic extract (25-30 mg/kg), hexane (10-50 mg/kg, i.p.) and ethylacetate (50 and 100 mg/kg i.p.) fractions produced significant inhibition (P<0.01) of the acetic acid-induced writhing in mice and suppressed formalin-induced licking response of animals in both phases of the test. In the hot plate assay the plant extract (100 mg/kg i.p.) increased pain threshold of mice. Pre-treatment of animals with naloxone (5 mg/kg i.p.) abolished the analgesic effect of the C. oxycantha in formalin and hot plate tests. C. oxycantha (50-200 mg/kg i.p.) produced marked anti-inflammatory effect in carrageenan-induced rat paw edema assay comparable to diclofenac. These findings suggest that C. oxycantha possesses central analgesic and peripheral anti-inflammatory properties, with analgesic effects associated with the opioid system.